Edited by: Hans-Uwe Simon
Possible therapeutic potential of a recombinant group 2 grass pollen allergen-specific antibody fragment
Article first published online: 20 NOV 2013
© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd
Volume 69, Issue 2, pages 261–264, February 2014
How to Cite
Possible therapeutic potential of a recombinant group 2 grass pollen allergen-specific antibody fragment. Allergy 2014; 69: 261–264., , , , .
- Issue published online: 27 JAN 2014
- Article first published online: 20 NOV 2013
- Manuscript Accepted: 2 OCT 2013
- Austria Science Fund (FWF), by the Christian Doppler Research Association, Biomay. Grant Numbers: P23318-B11, F4605, F4607, F4611
- grass pollen;
- recombinant single-chain fragment;
The induction of blocking IgG antibodies that compete with IgE for allergen binding is one important mechanism of allergen-specific immunotherapy. The application of blocking antibodies may be an alternative treatment strategy. A synthetic gene coding for a single-chain fragment (ScFv) specific for the major timothy grass pollen allergen Phl p 2 was inserted into plasmid pCANTAB 5 E, and the recombinant ScFv was expressed in Escherichia coli and purified by affinity chromatography. The ScFv was tested for allergen binding by ELISA, and its association and dissociation were measured by surface plasmon resonance (Biacore) technology. The ability of the ScFv to inhibit allergic patients' IgE binding to Phl p 2 and Phl p 2-induced basophil degranulation was studied by ELISA competition and basophil activation (CD203c) assays. We report the expression, purification, biochemical and immunological characterization of a monomeric single-chain fragment (ScFv) of human origin specific for the major timothy grass pollen allergen, Phl p 2. The Phl p 2-ScFv showed high affinity binding to the allergen and blocked the binding of allergic patients' polyclonal IgE to Phl p 2 up to 98%. Furthermore, it inhibited allergen-induced basophil activation. The Phl p 2-ScFv inhibited allergic patients' IgE binding to Phl p 2 as well as Phl p 2-induced basophil activation and might be useful for passive immunotherapy of grass pollen allergy.